Scaffold-based 3D cell culture models in cancer research

Abstract

Three-dimensional (3D) cell cultures have emerged as valuable tools in cancer research, offering significant advantages over traditional two-dimensional (2D) cell culture systems. In 3D cell cultures, cancer cells are grown in an environment that more closely mimics the 3D architecture and complexity of in vivo tumors. This approach has revolutionized cancer research by providing a more accurate representation of the tumor microenvironment (TME) and enabling the study of tumor behavior and response to therapies in a more physiologically relevant context. One of the key benefits of 3D cell culture in cancer research is the ability to recapitulate the complex interactions between cancer cells and their surrounding stroma. Tumors consist not only of cancer cells but also various other cell types, including stromal cells, immune cells, and blood vessels. These models bridge traditional 2D cell cultures and animal models, offering a cost-effective, scalable, and ethical alternative for preclinical research. As the field advances, 3D cell cultures are poised to play a pivotal role in understanding cancer biology and accelerating the development of effective anticancer therapies. This review article highlights the key advantages of 3D cell cultures, progress in the most common scaffold-based culturing techniques, pertinent literature on their applications in cancer research, and the ongoing challenges.

Keywords: Decellularized tissues; Extracellular matrix (ECM); Hydrogels; Microfluidics; Scaffolds; Three-dimensional (3D) cell culture.

Fig. 1

Characteristics of 2D and 3D cell cultures

Fig. 2

Classification of polymers used for fabricating polymer-based 3D cell culture scaffolds

Fig. 3

Common methods of hydrogel 3D cultures: A the dome technique: cells are mixed with temperature-sensitive hydrogels then seeded as droplets in the cell culture vessel, then carefully covered with media, B insert wells: media is added in the well whereas cell suspension (cell in hydrogel mix) is placed in the insert, then covered with another layer of media. Heterogeneous spheroids will form on the insert bottom, C gel-bottom support: the bottom of the well is covered with a thick layer of hydrogel, on top of which the cell suspension is placed, and D embedding technique: cells mixed with hydrogel are placed on the bottom and then covered with a layer of media to support spheroid growth in the matrix

Fig. 4

Preparation methods, characterization techniques, and sources of decellularized tissues used as scaffolds for 3D cell culture. SEM: scanning electron microscopy; AFM: atomic force microscopy; FTIR: Fourier-transform infrared spectroscopy

Fig. 5

illustration of the microfluidic chip used in 3D co-culture of human colorectal cancer cells (HT-29) and normal colorectal fibroblasts (CCD-18Co) in a collagen matrix. The chip comprised 4 units, each featuring 7 channels for cell loading or media fill. Cancer and fibroblast cells were loaded into channels 4 and 2 in the co-culture, while channels 1 and 3 were designated for media fill. A cell loading channel’s detailed structure and dimensions are illustrated at the bottom left. Figure adapted from [145]